1 Joint Department of Biomedical Engineering at University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, and North Carolina State University , Raleigh, North Carolina.
Tissue Eng Part A. 2014 Jan;20(1-2):67-78. doi: 10.1089/ten.TEA.2013.0006. Epub 2013 Sep 19.
We have shown that the uniaxial cyclic tensile strain of magnitude 10% promotes and enhances osteogenesis of human mesenchymal stem cells (hMSC) and human adipose-derived stem cells (hASC) from normal, nonosteoporotic donors. In the present study, MSC from osteoporotic donors were analyzed for changes in mRNA expression in response to 10% uniaxial tensile strain to identify potential mechanisms underlying the use of this mechanical loading paradigm for prevention and treatment of osteoporosis. Human MSC isolated from three female, postmenopausal osteoporotic donors were analyzed for their responses to mechanical loading using microarray analysis of over 47,000 gene probes. Human MSC were seeded in three-dimensional collagen type I constructs to mimic the organic extracellular matrix of bone and 10% uniaxial cyclic tensile strain was applied to promote osteogenesis. Seventy-nine genes were shown to be regulated within hMSC from osteoporotic donors in response to 10% cyclic tensile strain. Upregulation of six genes were further confirmed with real-time RT-PCR: jun D proto-oncogene (JUND) and plasminogen activator, urokinase receptor (PLAUR), two genes identified as potential key molecules from network analysis; phosphoinositide-3-kinase, catalytic, delta polypeptide (PIK3CD) and wingless-type MMTV integration site family, member 5B (WNT5B), two genes with known importance in bone biology; and, PDZ and LIM domain 4 (PDLIM4) and vascular endothelial growth factor A (VEGFA), two genes that we have previously shown are significantly regulated in hASC in response to this mechanical stimulus. Function analysis indicated that 10% cyclic tensile strain induced expression of genes associated with cell movement, cell proliferation, and tissue development, including development in musculoskeletal and cardiovascular systems. Our results demonstrate that hMSC from aged, osteoporotic donors are capable of enhanced osteogenic differentiation in response to 10% cyclic tensile strain with significant increases in the expression of genes associated with enhanced cell proliferation, musculoskeletal development, and angiogenesis. Surprisingly, cyclic tensile strain of magnitude 10% not only enhanced osteogenesis in hMSC from osteoporotic donors, but also enhanced expression of angiogenic factors. Better understanding and methodologies to promote osteogenesis in hMSC from elderly, osteoporotic donors may greatly facilitate achieving long-term success in bone regeneration and functional bone tissue engineering for this ever-growing patient population.
我们已经证明,幅度为 10%的单轴循环拉伸应变促进并增强了来自正常非骨质疏松供体的人骨髓间充质干细胞(hMSC)和人脂肪来源干细胞(hASC)的成骨作用。在本研究中,分析了来自骨质疏松供体的 MSC 对 10%单轴拉伸应变的 mRNA 表达变化,以确定使用这种机械加载范式预防和治疗骨质疏松症的潜在机制。使用超过 47000 个基因探针的微阵列分析,分析了从三名女性绝经后骨质疏松症供体中分离的人 MSC 对机械加载的反应。将人 MSC 接种在三维胶原 I 构建体中,以模拟骨的有机细胞外基质,并施加 10%的单轴循环拉伸应变以促进成骨作用。结果显示,来自骨质疏松供体的 hMSC 中有 79 个基因对 10%的循环拉伸应变有反应。实时 RT-PCR 进一步证实了六个基因的上调:原癌基因 jun D(JUND)和尿激酶型纤溶酶原激活物受体(PLAUR),这两个基因是网络分析中确定的潜在关键分子;磷酸肌醇 3-激酶,催化,δ 多肽(PIK3CD)和无翅型 MMV 整合位点家族成员 5B(WNT5B),这两个基因在骨生物学中具有已知的重要性;以及 PDZ 和 LIM 结构域 4(PDLIM4)和血管内皮生长因子 A(VEGFA),这两个基因我们之前已经表明,它们在响应这种机械刺激时在 hASC 中受到显著调节。功能分析表明,10%的循环拉伸应变诱导了与细胞运动、细胞增殖和组织发育相关的基因表达,包括肌肉骨骼和心血管系统的发育。我们的结果表明,来自老年骨质疏松症供体的 hMSC 能够在 10%的循环拉伸应变下增强成骨分化,与增强细胞增殖、肌肉骨骼发育和血管生成相关的基因表达显著增加。令人惊讶的是,幅度为 10%的循环拉伸应变不仅增强了来自骨质疏松症供体的 hMSC 的成骨作用,而且还增强了血管生成因子的表达。更好地了解和应用促进老年骨质疏松症供体 hMSC 成骨作用的方法,可能会极大地促进在这个不断增长的患者群体中实现骨再生和功能性骨组织工程的长期成功。